Rotenone: Difference between revisions
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:::: '''Preparation of 0.25 mM solution''' (dissolved in 99.9% ethanol) for '''0.5-mL O2k-chamber''': | :::: '''Preparation of 0.25 mM stock solution''' (dissolved in 99.9% ethanol) for '''0.5-mL O2k-chamber''': | ||
::::# Take 50 µL of the 1 mM stock solution | ::::# Take 50 µL of the 1 mM stock solution | ||
Revision as of 12:27, 22 October 2020
- high-resolution terminology - matching measurements at high-resolution
Rotenone
Description
Rotenone is an inhibitor of Complex I (CI) and thus inhibits NADH oxidation. It inhibits the transfer of electrons from iron-sulfur clusters in CI to ubiquinone via binding to the ubiquinone binding site of CI. See also Complex II-linked substrate state, S(Rot).
Abbreviation: Rot
Reference: Gnaiger 2014 MitoPathways, MiPNet09.12, MiPNet03.02
Application in HRR
- Rot: Rotenone (C23H22O6); Sigma R 8875, 1 g, store commercial stock at RT; FW = 394.42 (≥ 95%).
- Caution: Very toxic and light sensitive (store solution in a dark glass vial).
- Preparation of 1 mM stock solution (dissolved in 99.9% ethanol) for 2-mL O2k-chamber:
- Weigh 0.39 mg of rotenone and dissolve in 1 ml of ethanol absolute (99.9%).
- Divide into 100 µL aliquots (use dark glass vials).
- Store frozen at -20 °C.
- » O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume (2-mL O2k-chamber): 1 µL using a 10 µL Hamilton syringe.
- Final concentration: 0.5 µM.
- Preparation of 0.25 mM stock solution (dissolved in 99.9% ethanol) for 0.5-mL O2k-chamber:
- Take 50 µL of the 1 mM stock solution
- Add 150 µL of ethanol absolute (99.9%).
- » O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume (0.5-mL O2k-chamber): 1 µL using a 10 µL Hamilton syringe.
- Final concentration: 0.5 µM.
- Microalgae applications
- For the specific case of microalgal cells it is recommended to prepare higher concentration:
- Preparation of 5.5 mM stock solution (dissolved in 99.9% ethanol)
- Weigh 4.4 mg of rotenone and dissolve in 2 ml of ethanol absolute (99.9%).
- Divide into 200 µl aliquots (use dark glass vials).
- Store frozen at -20 °C.
- » O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume (2-mL O2k-chamber): 15 µL using a 25 µL Hamilton syringe.
- Titration volume (0.5-mL O2k-chamber): 3,75 µL using a 10 µL Hamilton syringe.
- Final concentration: 41 µM.
Inhibition of CI by rotenone
- Fato R, Bergamini C, Bortolus M, Maniero AL, Leoni S, Ohnishi T, Lenaz G (2009) Differential effects of mitochondrial Complex I inhibitors on production of reactive oxygen species. Biochim Biophys Acta 1787:384–92. >>PMC2724837
MitoPedia topics: Inhibitor
